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I am relatively new to RNA seq. I have done some mouse tissue RNA seq and thought mouse RNA seq would be much more difficult than bacterial RNA seq. But I am WRONG!! I have used zymo direct-zol RNA kit to extract RNA from bacteria and it gave very good reading on nanodrop. I know nanodrop reading does not really indicate any quality. I got this kind of reading on bioanalyzer and the RIN number is only 3 and 23S/16S is around 2. does anyone know what's the problem here? thank you very much!
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Love this - good data definitely starts from good input, and poor input can only give relatively poor data. I particularly like the mention of Nanodrop/absorbance based methods for quantification. It's such a toss up if you'll get an accurate reading or what amounts to a randomly generated number, and a lot of library/sequencing related issues can be traced back to poor quant.07-01-2026, 11:43 AM
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I’m not a sequencing expert. I’m a purification scientist who uses NGS to evaluate workflows my group develops. With this perspective, we think about the sample first and the NGS workflow second. The sequencer is an exceptionally honest reporter, but it can only report on what you give it, so whether you get clean, interpretable data from an NGS workflow is largely determined before you begin.
Here are nine questions we think about, in roughly the order they matter, before...